New therapeuhcally useful derivatives



United States Patent The invention relates to new therapeutically useful steroid compounds, that is to new derivatives of 4-hydroxy-l7a-methyl-testeosterone having the general formula:

wherein X=H, Br or is lacking when there is a 1,2 double bond,

Y=H when X=Br or is lacking, Cl when X=H or Br, or is lacking when there is a 6,7 double bond and when X=H or is lacking, and I R=H or Ac where Ac=acyl group derived from an acid containing not more than 9 carbon atoms,

and to processes of preparing them.

B. Camerino et al. have disclosed (in J. Am. Chem. Soc. 78, 1956, page 3540), a number of steroids of the androstane series (having either a chlorine or a hydroxy group in 4-position), which possess outstanding anabolic activity and low androgenic effect. Moreover, all of the products described in the publication are highly active when administered subcutaneously.

More recently, B. Camerino et al. have described (in British patent specification 848,288) 4-hydroXy-17amethyl-testosterone, which has a high anabolic activity and low androgenic effect, even when administered orally. Orally active anabolic substances are especially sought after in the clinic.

The new products of the invention possess oral anabolic potency which is superior to that of 4-hydroxy-17a-methyl-testosterone and therefore represent further real progress in the search for orally administnable anabolic substances which serve either to stimulate proteic anabolism, to improve the general condition, or to increase body weight.

In particular, compounds:

2.5-bromo-4-hydroxy-17a-methyl-testosterone, 2.5-bromo-4-hydroxy-l 7 a-methyl-testosterone-4-acetate, 1,2-dehydro-4-hydroxy-17 a-rnethyl-testosterone, 1,2-dehydro-4-hydroxy-17a-rne1thyl-testosterone-4-acetate, 6g-chloro-4-hydroxy-l7a-methyl-testosterone,

the invention provides the following new fig-chloro-4-hydroxy-17a-methyl-testosterone-4-acetate,

6,7 -dehydro-4-hydroxy-17u-methy1-testosterone, 6,7-dehydro-4-hydroxy-17ot-rnethyl-testosterone-4-acetate, 1,2: 6,7 -dehydro-4-hydroxyl7 a-methyl-testostlerone, 1,2:6,7 dehydro 4 hydroxy 17a methyl testosterone-4-acetate, and 2g-bromo-6g-chlorot-hydroxy-l7a-methyl-testosterone.

-It is understood that other 4-acyloxy products having the aforesaid general structural formula are included with- 351033522 Patented Sept. 10, 1963 in the scope of the present invention. The 4-esters can be derived from aliphatic or cycloaliphatic or aromatic acids containing not more than 9 carbon atoms, such as, for example, formate, acetate, propionate, butyrate, valerate, succinate, heptanoate, cyclopentylpropionate, cyclohexaneacetate, hexahydrobenzoate, benzoate, phenylpropionate and analogues.

The products of the present invention are prepared from 4-hydroxy-(or 4-acyl0xy )17 a methyl-testosterones described by B. Camerino et al. in the British patent specification 848,288.

The synthesis of the products of the invention is carried out substantially in accordance with the following schematic illustration:

N WU

3 l OR Cl (VII, R=H) (VI, R=H) \(VIIa, R=Ac) 7 (Via, R=Ae) OH W011,

The process illustrated comprises halogen-ating 4-hydroxy-(or 4-acyloxy-) 17 a-methyl-testosterone (I and Ia), either with bromine in the 2-position to yield compound II or IIa, or with chlorine in 6-position to yield compound IV or IVa. This is followed by dehydrohalogenation to give products which contain respectively a double bond in the 1,2-position (III or IIIa) or in the 6,7-position (V or Va), or two double bonds in the 1,2- and 6,7-positions (VI or VIa). Products VI and VIa may alternatively be obtained from compound I or Ia by dehydrogenation in the 1,2- and 6,7-positions. Esterification of the 4-hy-droxy group or hydrolysis of the 4-acyloxy group may be carried out between any steps in the synthesis.

A most surprising aspect of the process of the invention is that the halogenation with bromine occurs in the 2-position of the steroid nucleus, while the halogenation with chlorine occurs in the 6-position.

The formation of A and A derivatives of 4-hydroxy- (or 4-a cyloxy-) l7a-methy1-testosterones both by dehydrohalogenation of the corresponding 2-bromoor 6- ch-loro-derivatives and by dehydrogenation in the 1,2- and 6 ,7-positi0ns was quite unexpected in view of the presence of the hydroxy group in the 4-position, which may exist in a keto form, and in view of the hydroxy group in the 17-position whose elimination is known to occur very easily.

4-hydroxy-(or 4-acyloxy-)17a-methyl-testosterones (I or Ia) may be brorninated in the 2-position either with bromine or with N-bromo-succinimide in an organic halogenated solvent such as carbon tetrachloride, chloroform or methylene dichloride or in acetic acid, to produce 2.2-bromo-4-hydroxy-(or 4-acyloxy).l7a-methyl-testoster- V ones (II or IIa), or may be chlorinated in the 6-position either with chlorine or with N-chloro-succinim-ide in an organic halogenated solvent such as carbon tetrachloride, chloroform, or methylene dichloride or in acetic acid, to yield 65 chloro-4-hydroxy-(or 4-acyloxy) 17a methyltestosterones (IV or IVa).

Both by bromination in the 2-position of 65-chloroderivatives (IV and IVa) and by chlorination in the 6- position of 2&bromo derivatives (II and 1111), the 25- bromo-6fi-chloro derivatives (VII and VIIa) may be obtained.

1,2 dehydro-4-hydroxy-(o-r 4'acyloxy-) 17a methyltestosterones (III or IIIa) may be obtained either from the 2-bromo-derivatives (II and IIa) by dehydrobromination with lithium bromide and lithium carbonate in dimethylformamide under a nitrogen atmosphere at about 80110 C. for 10-30 hours, or from the starting compounds I or Ia by dehydrogenation in the 1,2-position with selenium dioxide in ttbutanol and optionally in the presence of acetic acid.

6,7-dehydro-4-hydroxy-(or 4 acyloxy)l7a methyltestosterones (V or Va) are prepared analogously either from the 6-chlor0-derivatives (IV or IVa) by dehydrochlorination with lithium chloride and lithium carbonate in dimethylformamide or from the starting compounds I or Ia with chloranil in a warm aromatic solvent such as toluene.

1,2: 6,7-dehydro-4-hydroxy-(or 4-acyloxy-) Not-methyltestosterones -(VI or VIa) are produced either from the corresponding 6,7-dehydro-derivatives (V or Va) by dehydrogenation in the 1,2-psition with selenium dioxide, or from the corresponding 1,2-dehydro-derivatives (III or IIIa) by dehydrogenation in the 6,7-position with chloranil. The same products (VI and VIa) may be prepared from the corresponding 2g-bromo-6g-chloroderivatives (VII or VIIa), b-y dehydrohalogenation with lithium chloride, lithium bromide and lithium carbonate in the presence of dimethylformamide.

The acylation in the 4-position of the new compounds of the present invention may be carried out with the chloride or the anhydride of an organic acid having not more than 9 carbon atoms optionally in the presence of tertiary amines such as pyridine, between any steps of the synthesis. The hydrolysis of the acyloxy group in the 4-position of the new compounds having the aforesaid general structural formula may be effected between any steps of the synthesis with alkali according to known procedures.

The compounds of the invention show unexpected anabolic activity and are particularly useful when orally administered. In comparison with the known 4-hydroxy- 17u-methyl-testosterone and its esters, the compounds of the invention are more useful therapeutically, as shown in the following examples.

The invention includes therapeutic compositions comprising one or more compounds of the invention with a pharmaceutically [acceptable carrier which may be either a solid or a liquid. The compositions may be prepared as elixirs, tablets, powders, pills, capsules or in other dosage forms which are particularly suitable for oral administration. Liquid diluents, such as water, are employed in a sterile condition for parenteral use. Solid diluents and/or excipients include starch, lactose, talc, stearic acid, magnesium stearate, pectins and others generally used for these purposes. The active compounds may be used without diluents or excipients by placing them in the usual capsules such as gelatin capsules or other containers. The percentage of active ingredient in the composition may be varied according to the particular therapeutical form. In general, the compositions of the invention should contain at least 1%, preferably 575% of active substance.

The invention is illustrated by the following preferred embodiments:

Example 1 2E-BROMO4-HYDROXY-l7a-METHYL-TESTOSTERONE (II) FROM (I)-P'ROCESS WITH BROMINE 3 g. of 4-hydroxyl17oz-methyl-testosterone (I) are dissolved in 30 cc. of anhydrous chloroform; the solution is cooled to 10 C. and 1.44 g. of bromine, dissolved in 15 cc. of anhydrous chloroform, are added with stirring. A quick absorption of bromine is observed. The chloroform solution is washed with water, 5% aqueous sodium bicarbonate, then with water to neutrality, dried over anhydrous sodium sulphate and evaporated to dryness under vacuum at a temperature under 35 C. The residue is crystallized twice from aqueous methanol.

1.6 g. of (II), melting at 102-104 C. with decomposition are obtained.

at 286 m (e 10,750)

Example 2 ZE-BROM O 4'HYDROXY '17a METHYL-TESTOSTERONE (11) FROM (I)PRO CESS WITH N-BROMOSUCCINI- MIDE Example 3 2g-BROM0-4HYDROXY-lYa-METHYL IESTOSTERONE- 11-ACET'A'IE (Ila) AczCOC'Ha FROM (II) 1 g. of product (11), dissolved in 8 g. of pyridine is acetylated in known manner with 1 g. of acetic anhydride at room temperature. The product (Ila) melting at C. (recrystallized from ether/petroleum ether) is obtained.

A223? at 251 u Example 4 l,Z DEHYDR0-4-HYDROXY-l7a-MZETHYL-TESTOSTERONE (III) FROM (II) To 2 g. of 2E-bromo-4-hydroxy-17a-methyl-testos- 'ing at 130132 C. is obtained.

terone (II), dissolved in 30 cc. of dimethylformamide, 2 g. of lithium bromide and 1.33 g. of lithium carbonate are added under nitrogen with stirring. The mixture is allowed to react at 105 C. for 20 hours. The solution is cooled and poured into 200 cc. of Water; the steriod is extracted with ethyl acetate, the extract is washed with water to neutrality, dried over anhydrous sodium sulphate and evaporated to dryness. The residue is crystallised from ether/petroleum ether. After 2 crystallizations 0.61 g. of (III) melting at 158-161" C. are ob tained.

rar at 305 a ($5,100)

rate at 244 my (@14 0), I in in chloroform) Example 1,2-DEHYDRO-4-HYDROXY-17a METHYL-TESTOSTERONE i-ACETAT-E (Illa) AczCOCHa FROM (III) 100 mg. of 1,2-dehydro-4-hydroxy-l7a-methyl-testosterone (III), dissolved in 0.7 cc. of pyridine, are reacted with 0.15 cc. of acetic anhydride at room temperature. After 18 hours the solution is diluted with water, the steroid is extracted with ethyl acetate, the organic extract is washed with aqueous 5% hydrochloric acid, then with water, with aqueous 5% sodium bicarbonate and finally with water to neutrality. The extract is dried over anhydrous sodium sulphate and evaporated to dryness under vacuum.

The residue is recrystallized from ether/petroleum ether. The product (Illa) meltggr at 246 mu (e= Example 6 1,2-DEHYDRO 4-HYDROXY-I'YwMETHYL-TESTOSTERONE- ii-ACETATE (IIIa) ACZCOCHs FROM (Ia) AczCOCHa '1 g. of 4-hydroxy-17a-methyhtestosterone-4-acetate (Ia) are reacted with 0.33 g. of selenium dioxide and 0.2 cc. of acetic acid in 20 cc. of t.butanol, at 70 C. under nitrogen with stirring. After 8 hours 0.33 g. of selenium dioxide are added and the reaction is prolonged for another 16 hours. The mixture is evaporated to dryness under vacuum and the residue is treated with ethyl acetate. The solution is washed with aqueous 7% ammonium sulphide, aqueous 7% ammonium hydroxide, water, aqueous 5% hydrochloric acid and finally with water to neutrality. The solution is dried over anhydrous sodium sulphate and evaporated to dryness under vacuum. The residue is recrystallized from ether/petroleum ether. After several recrystallizations the product (Illa) (Ac=COCH melting at 130-132 C. and identical with that produced as described in Example 5, is obtained.

Example 7 fig-CEtLOxlito-lrHYDROXY 17 a METHYL-TESTOSTERONE (I FROM (I)-PROCESS WITH CHLORINE- To a solution of 2 g. of 4-hydroxy-l7oc-methyl-testosterone (I) in 10 cc. of anhydrous chloroform, 0.45 g.

of chlorine dissolved in 3.4 cc. of anhydrous chloroform are added. The chloroform solution is washed with water, aqueous 5% sodium bicarbonate and with water X2212 at 282 m (e=11,300), M1 7;l:2 (0. 1% in chloroform) Example 8 6CHLORO4HYDROXY 17 a. METHYL-TESTOSTERONE (IV) FROM (I)PROCESS WITH N-CI-ILOROSUCCINI- MIDE 1.59 g. of 4-hydroxy-17a methyl-testosterone (I) are reacted with 0.67 g. of N-chlorosuccinimide in'35 cc. boiling chloroform for 30 minutes and under direct ilto neutrality, dried over anhydrous sodium sulphate and i ,200 cc. of water lumination. The mixture is cooled, filtered and the filtrate is evaporated to dryness at a temperature below 35 C. The residue is crystallized from ether, the product (IV), identical with that yielded as described in Example 7, is obtained.

Example 9 62-01113ORO 4-HYDROXY-17a-METHYL-TESTOSTERONE- it-ACETATE (Iva) Ac:C OCHa FROM (IV) 200 mg. of 6.5-chloro-4-hydroxy-l 7ocmethyl-testosterone (IV) are dissolved in 1 cc. of pyridine and reacted with 0.2 cc. of acetic anhydride at room temperature. After 18 hours the solution is diluted with Water, the steroid is extracted with ethyl acetate, the extract is washed with aqueous 5% hydrochloric acid, water, aqueous 5% sodium bicarbonate and finally with water to neutrality. The extract is dried over anhydrous sodium sulphate and evaporated under vacuum at a temperature below 35 C. The residue is crystallized from ether. The product (IVa) (Ac=COCH melting at 155-458 C. is obtained.

Example 10 6,7-DEHYDRO-4-HYDROXYJYwM'ETHYL-TESTOSTERONE (V) FROM (IV) 1.1 g. of 65-chloro-4-hydroxy-17a-rnethyl-testosterone (IV) dissolved in 26 cc. of dimethyltormamide are reacted with 1.70 g. of lithium chloride and 1.20 g. of lithium carbonate under nitrogen with stirring at C. for 20 hours. The solution is cooled and poured into The steroid is extracted with ethyl acetate, the extract is washed with water to neutrality, dried over anhydrous sodium sulphate, decolorized with active carbon and. concentrated to small volume. The residual solution is filtered, treated with ether and the product is filtered, washed. with ether and crystallized twice from ethyl acetate.

xglzganol t m Example 11 6 ,7-DEHYDRO-4-HYDROXY-1 "(a-METHYL-TE STO STERONE (Va) Ac: COCHs FROM (V) By acetylation of (V), dissolved in pyridine, with acetic anhydride, the 4-acetate (Va) (Ac=COCH melting at 130 C. is obtained.

53 21101 t 289 m (::2l,500)

Example 12 6;7-DEHYDRO-4-HYDROXY-17a-METHYL-TESTOSTERONE- 4ACETATE (Va) Ac: COCHs FROM (Ia) Ac=COCH3 xgm at 289 mu Example 13 1,2 6,7-DEHYDRO-4-HYDROXY 17a METHYL- TESTOS- TERONE- i-ACETATE (VIa) Ac: COCHa FROM (Va) ACZCOCHS By dehydrogenation, in known manner, of Va dissolved in t.butanoi with selenium dioxide and acetic acid, as described in Example 6, the product (VIa) Ac=COCH melting at 166-1169 C. is obtained.

xii 1211 at 225 mp. =12,800)

Age at 252 mu ima at 305 mp (e=1 7 Example 14 1,2 6,7-DEHYDRO-4-HYDROXY-17a-MIE'1HYLTESTOS- 'I'ERONE (VI) FROM (VIa) AczCOCHa Ethanol max.

at 232 m 11,800)

A225) at 348 m (e=8,400)

Example 15 2-BROMO 6=CHLORO 4-HYDROXY-1'7a-METHYL- TESTOSTERONE (VII) FROM (II) 1.20 g. of 25-bromo-4-hydroxy-17a-methyl-testosterone dissolved in 7.5 cc. of anhydrous chloroform are reacted with 212 mg. of chlorine dissolved in 2.1 cc. of anhydrous chloroform at 5 C. With stirring. A quick absorption of chlorine is observed. The chloroform solution is Washed with water, aqueous 5% sodium bicarbonate and finally with water to neutrality. The extract is dried over anhydrous sodium sulphate and evapo rated to dryness under vacuum at a temperature under 35 C. The residue is crystallized twice from ether. The product (VII) melting at 175185 C. with decomposition is obtained.

max.

at 293 m (e=9,500)

Example 16 2E-BROMO GE-CHLORO-4-HYDROXY-17a-METHYL- TESTOSTERONE (VII) FROM (IV) 2.07 g. of 6g-ch1oro-4-hydroxy-17a-methyl-testosterone dissolved in 15 cc. of anhydrous chloroform are reacted with 0.95 g. of bromine dissolved in 10 cc. of anhydrous chloroform at 5 C. with stirring. A quick absorption of bromine is observed. The chloroformic solution is washed with water, aqueous 5% sodium bicarbonate and finally with water to neutrality. The extract is dried over anhydrous sodium sulphate and evaporated to dryness under vacuum at a temperature under 35 C. The residue is treated with ether; the raw product (VII) melting at 155165 C. with decomposition is obtained. By crystallization from ether the M.P. rises up to 173-182 C. with decomposition.

Agg at 293 my $9,300)

Example 17 1,2: SW-DEHYDRO--HYDRO'XY-lla-METHYL- TESTOSTERONE (VI) FROM (VIII) 2 g. of 2i5-bromo-6g-chloro-4-hydroxy-l7a-methyl-testosterone, dissolved in 30 cc. of anhydrous dimethylformamide are reacted with 2.25 g. of lithium chloride, 2.25 g. of lithium bromide and 3.2 g. of lithium carbonate at 105 C. for 20 hours in a nitrogen atmosphere and with stirring. The solution is cooled and poured into 200 cc. of water; the steroid is extracted with ethyl acetate and the extract is washed with water, dried over anhydrous sodium sulphate and evaporated to dryness. The residue is crystallized many times from ether, and decolorized with active carbon. The product (VI) melting at 180-183" C. is obtained.

Ethunol max.

8 Example 18 TERONE-A-ACETATE (VItt) Ac: COCHz FROM (VI) mg. of 1,2:6,7-dehydro-4-hydroxy-l7a-methyl-testosterone dissolved in 1 cc. of pyridine are acetylated with 0.2 cc. of acetic anhydride at room temperature. After 18 hours the solution is diluted with water and the steroid is extracted with ether. The organic extract is washed with aqueous 5% hydrochloric acid, water, aqueous 5% sodium bicarbonate and finally with water to neutrality, dried over anhydrous sodium sulphate and concentrated to small volume. The product (VIa) Ac=COCH which after crystallization from ether melts at 166-169 C. is obtained.

xgtalgnol at m Aggy at 252 mp. (e=

gg e at 305 mu (e=12,000)

Example 19 2E-BROMO4-HYDROXY-l7a-METHYL TESTOSTERONE (II) FROM (I )BROMINATION IN ACETIC ACID 0.63 g. of bromine dissolved in 7.15 cc. of anhydrous acetic acid, are added, at 5 C. and with stirring to a solution of 1.25 g. of 4-hydroxy-l7a-methyl-testosterone (I) in 7 cc. of anhydrous acetic acid. The solution is poured into 70 cc. of water and the steroid is extracted wtih ether. The organic extract is washed with water, aqueous 5% sodium bicarbonate, water to neutrality, dried over anhydrous sodium sulphate and concentrated to small volume. The product (II), identical with that prepared as described in Examples 1 and 2, is obtained.

Example 20 2g-BR0M0 et-HYDEOXY-17a-METHYL-TESTOSTERONE (II) FROM (Ila) Ac=COCHa A mixture of 0.5 g. of 25-bromo-4-hydroxy-17a-methyltestosterone-4-acetate (Ila) Ac=COCH dissolved in 15 cc. of methanol and 0.5 g. of potassium bicarbonate dissolved in 5 cc. of Water, is allowed to react at room temperature for 18 hours. The solution is neutralized with acetic acid, poured into water, and the steroid is extracted with ether. The extract is Washed with water to neutrality, dried over anhydrous sodium sulphate and evaporated to dryness at a low temperature. The residue is crystallized from ether. The product (II), identical with that obtained as described in Examples 1, 2,.and 19, is obtained.

PHARMACOLOGICAL ACTIVITY The anabolic and androgenic activities of a number of products of the present invention are hereinafter reported in comparison with those of 17a-methyl-testosterone and 4-hydroxy-17a-methyl-testosterone. The anabolic (myotropic) and androgenic properties have been determined in male castrated rats weighing 30-40 g., according to the method of Hershberg et al. (Proc. Soc. Exp. Biol. and Med. 83, 1953, page The increase in weight of levator ani muscle has been considered as an expression of anabolic activity, while the increase in weight of prostate gland has been considered as an expression of androgenic activity. The reported data in the following Table 1 are referred to wet weights of the organs.

For comparing the anabolic and endrogenic activities of the tested steroids the following ratio was established:

Experimental levator ani weight minus control levator ani weight The steroids were administered orally.

= therapeutic index Tl.)

Table 1.--Anablic and androgenic activities (oral admzmstratlon) Steroid Doses Levator Prostate T.I.

(mg/day) ani, mg.

Castrated controls 8.6 9. 3 1 13.4 33. 7 0.18 2 17.7 43. 9 0.26 4 25. 5 73. 7' 0.26

0.5 22. 6 32.4 0.61 O.. 1 22.8 30.7 0.66 D0 2 30. 7 48. 1 0.57 2E-bromo4-hydroxy-17amethyl-testosteronc(11) 0.5 19.2 22.0 0. 83 1,2-dehydro-4-hydroxy-17amethyl-testosterone(III) 0.25 14.8 19.2 0.63 D0 0.5 24. 7 21. 3 1. 34 6E-cl1loro-4-hyclroxy-17amethyl-testosterone (IV) 0.25 19. 5 19. 9 1.02 D0 0.5 21.6 24. 8 0.83 Do 1 25. 3 34. 0 0. G8 6E-chloro-4-hydroxy-17a- 0. 5 17. 6 21. 8 0. 72

methyl-testosteronei-acetate (IVa) Ac=CJOCH 1 21.9 26.4 0.77

The data of Table 1 show that the new products of the present invention, orally administered, possess a high myotropic activity, much higher than that of 17a-methyltestosterone, about equal to that of 4-hYdIOXY-17oc-Ille'tl1Yltestosterone, while their androgenic activity is much lower than those of the same comparison products.

Therefore the therapeutic index (T.I.) of the new products of the present invention is decidedly superior to that of 17u-methyl-testosterone and better than that of 4-hydroxy-17a-methyl-testosterone.

The relative anabolic potencies, determined at two dosage levels are reported in Table 2, with respect to our compounds 11, III, IV, and IVa.

The meaning of the undulated linkage of X and Y in 2- and 6-positions of the general formula (Greek script letter .5 (Xi)) is that said linkage is indeterminate.

We claim:

1. A derivative of 4-hydroxy-17a-methy1-testosterone, taken from the group consisting of those having one of the following formulae:

and

OH LH in which R is taken from the group consisting of H and Ac, the Ac radical being an acyl radical of the formula R CO in which R is a hydrocarbon radical having not more than nine carbon atoms.

2. A compound having the formula:

CH3 CH3 1 0H References Cited in the file of this patent UNITED STATES PATENTS 2,705,237 Djerassi et al. Mar. 29, 1955 2,762,818 Levy et a1. Sept. 11, 1956 2,837,464 Nobile June 3, 1958 2,882,282 Agnello et al Aug. 14, 1959 2,923,721 Joly et a1. .'Feb. 2, 1960 OTHER REFERENCES Ringold et al.: J. Org. Chem. 21, 239 (1956). 

1. A DERIVATIVE OF 4-HYDROXY-17A-METHYL-TESTOSTERONE, TAKEN FROM THE GROUP CONSISTING OF THOSE HAVING ONE OF THE FOLLOWNG FORMULAE: 